This method confirms the presence of the tranquilizer azaperone and its metabolite azaperol in swine liver tissue using gas chromatography/mass spectrometry (GC/MS). Swine liver is ground and extracted using acetonitrile and sodium chloride buffer. Solid phase extraction is used to isolate azaperone and azaperol, which are then eluted and analyzed by GC/MS. The method was validated by analyzing fortified tissue samples containing 10 ppb of both compounds and incurred tissue from pigs dosed with azaperone, confirming the presence of the parent drug and metabolite.
The automation of sample preparation has become an increasingly important component for reproducible and operator-independent experiments. This work outlines novel strategies that are being utilized for automated online and offline sample preparation to achieve specific goals, such as a host of applications including targeting post-translationally modified proteins, non-tryptic peptides, and intact proteins.
The presentation describes the automated process of the system and present a number of applications from sample matrices such as food, polymers, and pharmaceuticals to show the utility of the system.
Most commercial fermented milks showed moderate ACE-inhibitory activity, though some exceptions had higher activity that could relate to the milk origin. Two fermented milks were subjected to simulated gastrointestinal digestion, which increased their ACE-inhibitory activity, suggesting physiological digestion promotes formation of active peptides. Peptides generated from one product during digestion were sequenced, and most formed after 30 minutes of pancreatic enzyme incubation, indicating digestion facilitates peptide release from proteins in these fermented products.
The presence of Per- and Polyfluorinated Alkyl Substances (PFAS) in drinking water is being thoroughly studied due to the persistence of these compounds in the environment and their potential health effects. However, there is limited knowledge about the occurrence of these chemicals in bottled water, despite the increasing concerns about PFAS in the food supply. This poster shows results from a fast and simple direct injection method similar to draft EPA method 8237, using the Shimadzu triple quad LCMS-8050 to analyze seven commercially available samples of bottled water for 24 PFAS.
Analysis of Aflatoxins in Pet Food by UHPLC Using PDA and Fluorescence DetectionPerkinElmer, Inc.
Commercially prepared pet foods are easy and economical ways to fulfill the nutritional requirements for pets. Dry pet food is produced with grains and cereal by-products rejected for human consumption. The contamination of these by-products, with toxigenic fungal metabolites called mycotoxins, pose a serious health threat to pets.
Aflatoxins, some of the most carcinogenic mycotoxins known, are classified as B1, B2, G1, and G2. Several aflatoxin outbreaks in commercial pet foods have been reported in the past few years. Symptoms from aflatoxin exposure include lethargy, anorexia, jaundice, and intravascular coagulation, the severity often varying based upon a pet’s breed, species, age, dose, length of exposure, and nutritional status. Even if affecting only a small percentage of commercial pet foods, problems with pet food safety impact the entire pet food industry due to recalls and loss of consumer loyalty. Such experiences have reaffirmed the need for commercial pet food manufacturers to devote extensive resources documenting product quality.
Mycotoxins are strictly regulated around the world because of their strong carcinogenic effects. A simple and reliable method to analyze mycotoxins is required to ensure food safety. The current methods require time-consuming sample pretreatment. This presentation reports on a fully automated online sample extraction and analysis of mycotoxins in foods by online SFE-SFC-MS.
This document summarizes research identifying compounds from Adenocarpus cincinnatus that modulate GABAA receptors. Using HPLC-based activity profiling, 15 flavonoid and isoflavonoid derivatives were isolated from the plant, including 8 new compounds. Key findings include:
1) Isoflavone 11 and pterocarpans 2 and 8 showed promising GABAA receptor modulatory activity with EC50 values between 2.8-18.8 μM.
2) Pterocarpans were identified for the first time as a new scaffold for GABAA receptor modulators.
3) The isolated compounds' structures were elucidated using NMR and ECD spectroscopy, with many possessing a
This document summarizes a presentation on biological medicines and monoclonal antibody therapeutics. It discusses regulatory guidelines for biosimilars in Korea and ASEAN countries. The presentation provided an overview of global biosimilar guidelines, noting that the EU implemented the world's first well-organized biosimilar legislation and guidelines. It also noted that Korea has issued biosimilar guidelines since 2009, and Malaysia and Singapore are leading implementation of biosimilar guidelines in ASEAN countries.
The automation of sample preparation has become an increasingly important component for reproducible and operator-independent experiments. This work outlines novel strategies that are being utilized for automated online and offline sample preparation to achieve specific goals, such as a host of applications including targeting post-translationally modified proteins, non-tryptic peptides, and intact proteins.
The presentation describes the automated process of the system and present a number of applications from sample matrices such as food, polymers, and pharmaceuticals to show the utility of the system.
Most commercial fermented milks showed moderate ACE-inhibitory activity, though some exceptions had higher activity that could relate to the milk origin. Two fermented milks were subjected to simulated gastrointestinal digestion, which increased their ACE-inhibitory activity, suggesting physiological digestion promotes formation of active peptides. Peptides generated from one product during digestion were sequenced, and most formed after 30 minutes of pancreatic enzyme incubation, indicating digestion facilitates peptide release from proteins in these fermented products.
The presence of Per- and Polyfluorinated Alkyl Substances (PFAS) in drinking water is being thoroughly studied due to the persistence of these compounds in the environment and their potential health effects. However, there is limited knowledge about the occurrence of these chemicals in bottled water, despite the increasing concerns about PFAS in the food supply. This poster shows results from a fast and simple direct injection method similar to draft EPA method 8237, using the Shimadzu triple quad LCMS-8050 to analyze seven commercially available samples of bottled water for 24 PFAS.
Analysis of Aflatoxins in Pet Food by UHPLC Using PDA and Fluorescence DetectionPerkinElmer, Inc.
Commercially prepared pet foods are easy and economical ways to fulfill the nutritional requirements for pets. Dry pet food is produced with grains and cereal by-products rejected for human consumption. The contamination of these by-products, with toxigenic fungal metabolites called mycotoxins, pose a serious health threat to pets.
Aflatoxins, some of the most carcinogenic mycotoxins known, are classified as B1, B2, G1, and G2. Several aflatoxin outbreaks in commercial pet foods have been reported in the past few years. Symptoms from aflatoxin exposure include lethargy, anorexia, jaundice, and intravascular coagulation, the severity often varying based upon a pet’s breed, species, age, dose, length of exposure, and nutritional status. Even if affecting only a small percentage of commercial pet foods, problems with pet food safety impact the entire pet food industry due to recalls and loss of consumer loyalty. Such experiences have reaffirmed the need for commercial pet food manufacturers to devote extensive resources documenting product quality.
Mycotoxins are strictly regulated around the world because of their strong carcinogenic effects. A simple and reliable method to analyze mycotoxins is required to ensure food safety. The current methods require time-consuming sample pretreatment. This presentation reports on a fully automated online sample extraction and analysis of mycotoxins in foods by online SFE-SFC-MS.
This document summarizes research identifying compounds from Adenocarpus cincinnatus that modulate GABAA receptors. Using HPLC-based activity profiling, 15 flavonoid and isoflavonoid derivatives were isolated from the plant, including 8 new compounds. Key findings include:
1) Isoflavone 11 and pterocarpans 2 and 8 showed promising GABAA receptor modulatory activity with EC50 values between 2.8-18.8 μM.
2) Pterocarpans were identified for the first time as a new scaffold for GABAA receptor modulators.
3) The isolated compounds' structures were elucidated using NMR and ECD spectroscopy, with many possessing a
This document summarizes a presentation on biological medicines and monoclonal antibody therapeutics. It discusses regulatory guidelines for biosimilars in Korea and ASEAN countries. The presentation provided an overview of global biosimilar guidelines, noting that the EU implemented the world's first well-organized biosimilar legislation and guidelines. It also noted that Korea has issued biosimilar guidelines since 2009, and Malaysia and Singapore are leading implementation of biosimilar guidelines in ASEAN countries.
CYP2A6_HPLC_PK_2015 New Simple Method for Coumarin in Liver Cytochrome of RatsWael Ebied
This document describes the development and validation of a new liquid chromatography method for determining coumarin and its 7-hydroxy metabolite as a marker of cytochrome P450 2A6 activity in rats. The method uses a C18 column and isocratic mobile phase to separate coumarin, 7-hydroxycoumarin, and an internal standard. The method was validated and showed good linearity, precision, and accuracy. This validated method was then applied to study the effect of resveratrol, sulforaphane, and thymoquinone on hepatic CYP2A6 activity in spontaneously hypertensive rats.
This document summarizes a study that screened eight Monascus strains for their ability to produce lovastatin, an alternative to statin drugs. Strain M. purpureus Went (JCM 6934) produced the highest amount of lovastatin at 84.85 ppm. Angkak rice fermented with the local Philippine strain M. purpureus UPLB-MNH-MCC 2108 was found to also contain lovastatin. This showed that lovastatin is a characteristic component of angkak rice. A preliminary study found very low levels of the toxin citrinin in cookies made with angkak rice, showing it is safe for human consumption.
This document summarizes various experiments analyzing mitochondrial ribosome interacting proteins and overexpression of EF-Tu and EF-G proteins in E. coli. Figure 1 shows a SDS-PAGE gel with labeled mitochondrial ribosome subunit samples. The second section provides criteria for database analysis of mitochondrial ribosome associated proteins. Figures 2-4 show SDS-PAGE gels and western blots analyzing overexpression of EF-Tu and purification of EF-Tu and EF-G. Figures 7-8 again examine overexpression of EF-Tu and EF-G using SDS-PAGE and western blot.
This document describes the characterization and purification of lactate dehydrogenase (LDH) from bovine myocardium tissue. The authors developed quantitative assays to analyze and purify LDH activity extracted from the tissue. Through a series of fractionation and chromatography steps, including ammonium sulfate fractionation, ion exchange chromatography, and affinity chromatography, the authors were able to purify LDH from the myocardial extract into near homogeneity. They also determined physical and enzymatic characteristics of the purified LDH through assays and SDS-PAGE gel electrophoresis analysis.
Glutathione S-transferase enzymes (GSTs) play central roles in phase II detoxification of both xenobiotics and endogenous compounds in almost all living organisms. The enzyme was extracted and partially purified from wheat leaves through a procedure including ammonium sulfate fractionation followed by dialysis and gel filtration chromatography. These procedures yielded a 7.14-fold purification with 71% recovery. Optimum activity conditions-pH, temperature and ionic strength-of the enzyme were determined. Its some kinetic properties such as Vmax, KM, and kcat were calculated for GSH and CDNB substrates. The kcat/KM values of the enzyme were 603.5 for GSH and 385.3 for CDNB. The native molecular weight of the enzyme was estimated to be 52 kDa based on its mobility in gel filtration column.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
The document describes experiments to express recombinant human factor IX coagulation factor in E. coli cells. Key steps included amplifying the factor IX gene via PCR, inserting the gene into a plasmid vector, transforming E. coli cells with the recombinant plasmid, and quantifying expression levels using qPCR and ELISA. Agarose gel electrophoresis confirmed successful PCR and plasmid digestion. However, individually performed blue/white screening and expression assays did not yield expected results, possibly due to technical issues. The goals were to develop skills in DNA manipulation and analysis for therapeutic protein production applications like treating hemophilia B.
This document summarizes experiments testing the antimicrobial activity of crude probiotic bacterial lysates and powder lysates using BacT/ALERT bottles and a BacT/ALERT system. Several sample sets were prepared with varying concentrations of lysates and inoculated with S. aureus. The time taken for bacterial growth detection in each bottle was recorded. For the lower lysate concentrations, growth times were similar to the controls. Higher concentrations of 2g/2ml of the powder and crude lysates showed delays in detection time, indicating antimicrobial activity. The crude lysates showed greater variability in detection times between samples than the powder lysates. The BacT/ALERT system was able to objectively measure antimicrobial effects compared to
This document describes the design, fabrication, and evaluation of a novel sustained release drug delivery system using rosuvastatin pharmacosomes. Rosuvastatin was complexed with lecithin phospholipids to form pharmacosomes via a hand shaking method. The pharmacosomes showed improved solubility, drug loading efficiency, and sustained release properties compared to pure rosuvastatin. In vivo studies in rats demonstrated the pharmacosomes more effectively lowered cholesterol and triglyceride levels over longer periods compared to standard rosuvastatin treatment. Thus, the rosuvastatin pharmacosomes represent a promising sustained release drug delivery system with benefits like enhanced bioavailability and reduced dosing.
This document discusses clearance concepts in pharmacokinetics. It defines clearance as the proportionality constant relating the rate of drug elimination to its concentration in blood or plasma. Clearance depends on blood flow and the extraction ratio of the clearing organ. The two main types of clearance are hepatic clearance and renal clearance. Renal clearance can be estimated by measuring creatinine clearance. The document provides equations for calculating clearance, area under the curve, and creatinine clearance and adjusting drug doses based on renal function.
The document describes a human anti-hemophilic vaccine produced from human plasma. It discusses the collection of plasma from healthy donors, the production process which involves precipitation and purification using column chromatography, and the assay used to determine potency by comparing clotting times. The production involves testing donor blood to ensure safety, separating plasma from blood within 12 hours, and using an ion exchange column to capture factor VIII. The purified fraction is tested for activity and virus inactivation is performed before lyophilization.
Method Development and Validation-Solid Phase Extraction-Ultra Performance Li...Partha Ray
This method validates a solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry (SPE-UPLC-MS/MS) method for quantifying the antibiotic pirlimycin in bovine feces and urine. The method was sensitive, accurate, and precise for quantifying pirlimycin at low nanogram per gram and nanogram per milliliter levels. The method was applied to samples from dairy cows administered pirlimycin intramammarily, detecting pirlimycin in feces at 40.5-287 ng/g and in urine at 46.1-254 ng/mL over 120 hours. This validated method can be used to study the environmental impact of antibiotic excretion from livestock
A Method for the Quantification of Ethanol Content in Consumable Fruit Juices...PerkinElmer, Inc.
Production of alcohol has been long established in society with many styles that take advantage of the metabolism of sugars into ethanol. While the production of ethanol is desirable for alcoholic beverages, it is undesirable for other beverages which contain sugars that do not wish to be sold as an alcoholic beverage. Such sugar metabolism is naturally occurring and is well understood to happen in raw fruit as well as processed juice and can vary by type, variety and maturation in the growing season. A new application has been developed in the accurate determination of ethanol content in samples of these products utilizing the PerkinElmer® TurboMatrix™ headspace (HS) autosampler for better reproducible results.
This document summarizes research on the purification, characterization, and antifungal activity of a chitinase enzyme from Streptomyces venezuelae P10. Key findings include:
1) The chitinase was purified using ammonium sulfate precipitation, chitin affinity chromatography, and DEAE-cellulose anion exchange chromatography, yielding a purified enzyme with a molecular weight of 66 kDa.
2) The purified chitinase demonstrated optimal activity at 35°C and pH 6-8 and showed antifungal activity against phytopathogens such as Aspergillus niger and Alternaria alternate.
3) Thin layer chromatography analysis identified the chitin
This presentation reports on the development of a GC FID method to accurately quantify ethanol and IPA concentrations in two hand sanitizer samples. By using nitrogen as the carrier gas, this method is cost-effective and ensures the product compliance with CDC and USP guidelines and regulations.
This document evaluates the use of hydrogen (H2) carrier gas as an alternative to helium for the analysis of haloacetic acids (HAAs) according to EPA Method 552.3. Chromatograms of HAA standards using H2 were nearly identical to those using helium. Calibration curves for HAAs using H2 had coefficients of determination over 0.995. Accuracy and repeatability results using H2 met EPA requirements. Using H2 instead of helium reduces analysis costs by 2.69 to 6.15 times depending on gas purity, making H2 a suitable alternative carrier gas.
This document describes a fully automated method for analyzing 25-OH-Vitamin D3/D2 and 3-Epi-25-OH Vitamin D3/D2 in serum using LC-MS/MS. The method utilizes protein precipitation, solid phase extraction, and gradient elution on an Ascentis Express F5 column with MS/MS detection. Validation results show good linearity, accuracy, and precision for quantifying both the active and inactive vitamin D metabolites and their epimers. The fully automated system is capable of processing 96 samples in under 17 hours.
El documento habla sobre la gran cantidad de basura que se encuentra en las montañas y parajes naturales debido a los domingueros que realizan excursiones en vehículo y dejan sus desperdicios atrás. A pesar de llegar en coche, lo que les facilitaría recoger la basura, muchos la dejan tirada creyendo erróneamente que al esconderla ensucian menos. El documento pide a todos ser más cívicos y no dejar huella de contaminación en la naturaleza.
This document discusses My Vision Express, an electronic health record and practice management system used by over 12,000 eye care professionals worldwide. It is used by both small single-person practices and large retail chains with hundreds of locations. Key features of My Vision Express include patient management, insurance processing, inventory management, electronic health records, and a patient portal. The system aims to streamline workflows and improve efficiency for eye care practices.
How employers can take control of unmanaged chronic disease health costs, and drive productivity across the whole workforce through better eyecare integrated into benefits packages
CYP2A6_HPLC_PK_2015 New Simple Method for Coumarin in Liver Cytochrome of RatsWael Ebied
This document describes the development and validation of a new liquid chromatography method for determining coumarin and its 7-hydroxy metabolite as a marker of cytochrome P450 2A6 activity in rats. The method uses a C18 column and isocratic mobile phase to separate coumarin, 7-hydroxycoumarin, and an internal standard. The method was validated and showed good linearity, precision, and accuracy. This validated method was then applied to study the effect of resveratrol, sulforaphane, and thymoquinone on hepatic CYP2A6 activity in spontaneously hypertensive rats.
This document summarizes a study that screened eight Monascus strains for their ability to produce lovastatin, an alternative to statin drugs. Strain M. purpureus Went (JCM 6934) produced the highest amount of lovastatin at 84.85 ppm. Angkak rice fermented with the local Philippine strain M. purpureus UPLB-MNH-MCC 2108 was found to also contain lovastatin. This showed that lovastatin is a characteristic component of angkak rice. A preliminary study found very low levels of the toxin citrinin in cookies made with angkak rice, showing it is safe for human consumption.
This document summarizes various experiments analyzing mitochondrial ribosome interacting proteins and overexpression of EF-Tu and EF-G proteins in E. coli. Figure 1 shows a SDS-PAGE gel with labeled mitochondrial ribosome subunit samples. The second section provides criteria for database analysis of mitochondrial ribosome associated proteins. Figures 2-4 show SDS-PAGE gels and western blots analyzing overexpression of EF-Tu and purification of EF-Tu and EF-G. Figures 7-8 again examine overexpression of EF-Tu and EF-G using SDS-PAGE and western blot.
This document describes the characterization and purification of lactate dehydrogenase (LDH) from bovine myocardium tissue. The authors developed quantitative assays to analyze and purify LDH activity extracted from the tissue. Through a series of fractionation and chromatography steps, including ammonium sulfate fractionation, ion exchange chromatography, and affinity chromatography, the authors were able to purify LDH from the myocardial extract into near homogeneity. They also determined physical and enzymatic characteristics of the purified LDH through assays and SDS-PAGE gel electrophoresis analysis.
Glutathione S-transferase enzymes (GSTs) play central roles in phase II detoxification of both xenobiotics and endogenous compounds in almost all living organisms. The enzyme was extracted and partially purified from wheat leaves through a procedure including ammonium sulfate fractionation followed by dialysis and gel filtration chromatography. These procedures yielded a 7.14-fold purification with 71% recovery. Optimum activity conditions-pH, temperature and ionic strength-of the enzyme were determined. Its some kinetic properties such as Vmax, KM, and kcat were calculated for GSH and CDNB substrates. The kcat/KM values of the enzyme were 603.5 for GSH and 385.3 for CDNB. The native molecular weight of the enzyme was estimated to be 52 kDa based on its mobility in gel filtration column.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
The document describes experiments to express recombinant human factor IX coagulation factor in E. coli cells. Key steps included amplifying the factor IX gene via PCR, inserting the gene into a plasmid vector, transforming E. coli cells with the recombinant plasmid, and quantifying expression levels using qPCR and ELISA. Agarose gel electrophoresis confirmed successful PCR and plasmid digestion. However, individually performed blue/white screening and expression assays did not yield expected results, possibly due to technical issues. The goals were to develop skills in DNA manipulation and analysis for therapeutic protein production applications like treating hemophilia B.
This document summarizes experiments testing the antimicrobial activity of crude probiotic bacterial lysates and powder lysates using BacT/ALERT bottles and a BacT/ALERT system. Several sample sets were prepared with varying concentrations of lysates and inoculated with S. aureus. The time taken for bacterial growth detection in each bottle was recorded. For the lower lysate concentrations, growth times were similar to the controls. Higher concentrations of 2g/2ml of the powder and crude lysates showed delays in detection time, indicating antimicrobial activity. The crude lysates showed greater variability in detection times between samples than the powder lysates. The BacT/ALERT system was able to objectively measure antimicrobial effects compared to
This document describes the design, fabrication, and evaluation of a novel sustained release drug delivery system using rosuvastatin pharmacosomes. Rosuvastatin was complexed with lecithin phospholipids to form pharmacosomes via a hand shaking method. The pharmacosomes showed improved solubility, drug loading efficiency, and sustained release properties compared to pure rosuvastatin. In vivo studies in rats demonstrated the pharmacosomes more effectively lowered cholesterol and triglyceride levels over longer periods compared to standard rosuvastatin treatment. Thus, the rosuvastatin pharmacosomes represent a promising sustained release drug delivery system with benefits like enhanced bioavailability and reduced dosing.
This document discusses clearance concepts in pharmacokinetics. It defines clearance as the proportionality constant relating the rate of drug elimination to its concentration in blood or plasma. Clearance depends on blood flow and the extraction ratio of the clearing organ. The two main types of clearance are hepatic clearance and renal clearance. Renal clearance can be estimated by measuring creatinine clearance. The document provides equations for calculating clearance, area under the curve, and creatinine clearance and adjusting drug doses based on renal function.
The document describes a human anti-hemophilic vaccine produced from human plasma. It discusses the collection of plasma from healthy donors, the production process which involves precipitation and purification using column chromatography, and the assay used to determine potency by comparing clotting times. The production involves testing donor blood to ensure safety, separating plasma from blood within 12 hours, and using an ion exchange column to capture factor VIII. The purified fraction is tested for activity and virus inactivation is performed before lyophilization.
Method Development and Validation-Solid Phase Extraction-Ultra Performance Li...Partha Ray
This method validates a solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry (SPE-UPLC-MS/MS) method for quantifying the antibiotic pirlimycin in bovine feces and urine. The method was sensitive, accurate, and precise for quantifying pirlimycin at low nanogram per gram and nanogram per milliliter levels. The method was applied to samples from dairy cows administered pirlimycin intramammarily, detecting pirlimycin in feces at 40.5-287 ng/g and in urine at 46.1-254 ng/mL over 120 hours. This validated method can be used to study the environmental impact of antibiotic excretion from livestock
A Method for the Quantification of Ethanol Content in Consumable Fruit Juices...PerkinElmer, Inc.
Production of alcohol has been long established in society with many styles that take advantage of the metabolism of sugars into ethanol. While the production of ethanol is desirable for alcoholic beverages, it is undesirable for other beverages which contain sugars that do not wish to be sold as an alcoholic beverage. Such sugar metabolism is naturally occurring and is well understood to happen in raw fruit as well as processed juice and can vary by type, variety and maturation in the growing season. A new application has been developed in the accurate determination of ethanol content in samples of these products utilizing the PerkinElmer® TurboMatrix™ headspace (HS) autosampler for better reproducible results.
This document summarizes research on the purification, characterization, and antifungal activity of a chitinase enzyme from Streptomyces venezuelae P10. Key findings include:
1) The chitinase was purified using ammonium sulfate precipitation, chitin affinity chromatography, and DEAE-cellulose anion exchange chromatography, yielding a purified enzyme with a molecular weight of 66 kDa.
2) The purified chitinase demonstrated optimal activity at 35°C and pH 6-8 and showed antifungal activity against phytopathogens such as Aspergillus niger and Alternaria alternate.
3) Thin layer chromatography analysis identified the chitin
This presentation reports on the development of a GC FID method to accurately quantify ethanol and IPA concentrations in two hand sanitizer samples. By using nitrogen as the carrier gas, this method is cost-effective and ensures the product compliance with CDC and USP guidelines and regulations.
This document evaluates the use of hydrogen (H2) carrier gas as an alternative to helium for the analysis of haloacetic acids (HAAs) according to EPA Method 552.3. Chromatograms of HAA standards using H2 were nearly identical to those using helium. Calibration curves for HAAs using H2 had coefficients of determination over 0.995. Accuracy and repeatability results using H2 met EPA requirements. Using H2 instead of helium reduces analysis costs by 2.69 to 6.15 times depending on gas purity, making H2 a suitable alternative carrier gas.
This document describes a fully automated method for analyzing 25-OH-Vitamin D3/D2 and 3-Epi-25-OH Vitamin D3/D2 in serum using LC-MS/MS. The method utilizes protein precipitation, solid phase extraction, and gradient elution on an Ascentis Express F5 column with MS/MS detection. Validation results show good linearity, accuracy, and precision for quantifying both the active and inactive vitamin D metabolites and their epimers. The fully automated system is capable of processing 96 samples in under 17 hours.
El documento habla sobre la gran cantidad de basura que se encuentra en las montañas y parajes naturales debido a los domingueros que realizan excursiones en vehículo y dejan sus desperdicios atrás. A pesar de llegar en coche, lo que les facilitaría recoger la basura, muchos la dejan tirada creyendo erróneamente que al esconderla ensucian menos. El documento pide a todos ser más cívicos y no dejar huella de contaminación en la naturaleza.
This document discusses My Vision Express, an electronic health record and practice management system used by over 12,000 eye care professionals worldwide. It is used by both small single-person practices and large retail chains with hundreds of locations. Key features of My Vision Express include patient management, insurance processing, inventory management, electronic health records, and a patient portal. The system aims to streamline workflows and improve efficiency for eye care practices.
How employers can take control of unmanaged chronic disease health costs, and drive productivity across the whole workforce through better eyecare integrated into benefits packages
Los periféricos de entrada como teclado, ratón y micrófono permiten introducir datos al sistema, mientras que los periféricos de salida como monitor, impresora y altavoces permiten mostrar u obtener la salida de datos. Algunos periféricos como módem y multifunción permiten tanto la entrada como la salida de datos. Los sistemas de almacenamiento como discos duros, disquetes y CD-ROM/DVD-ROM almacenan datos de forma permanente o temporal.
BlackRock is the largest asset management firm globally, managing over $4.77 trillion in assets. It offers a wide range of products including mutual funds, ETFs, and advisory services. BlackRock has a history of acquisitions that have expanded its capabilities. It faces competition from other large asset managers but maintains strength in its brand, scale, and risk management platform called Aladdin. Both opportunities in emerging markets and threats from regulation and economic downturns could impact BlackRock's future performance.
This document provides tips for eye health and protecting vision. It recommends sitting farther from computer screens to avoid eye strain from lights. Taking regular breaks to blink more and adjusting screen colors can help. Seeing an optician regularly for checkups and treatments is also suggested, as is wearing UV-protected glasses, eating a vitamin-rich diet, avoiding smoking, and drinking alcohol in moderation.
El documento describe los controles comunes de Visual Basic, incluyendo etiquetas, cuadros de texto y botones de comando. Detalla las propiedades, eventos y métodos asociados con el control de etiqueta, que permite mostrar texto estático. También presenta un ejemplo de cómo cambiar los estilos de fuente de una etiqueta en respuesta a eventos de botón.
El documento describe un taller sobre el desarrollo de aplicaciones móviles con Xamarin. En el taller, los participantes crearán una aplicación meteorológica con información real utilizando conceptos como la estructura del proyecto, MVVM, diseño de interfaces y navegación. El taller se llevará a cabo de forma gradual a lo largo de 2,5 horas con explicaciones sobre Xamarin y demostraciones prácticas de cada parte de la aplicación.
En esta sesión veremos como adaptar nuestras aplicaciones para otorgar la mejor experiencia posible en teléfonos y tabletas. Como adaptar vistas, tener vistas específicas, adaptar navegación o detectar DPIs y tamaño de pantalla serán algunos de los puntos que veremos.
Xamarin.Forms es un framework que nos añade una capa de abstracción permitiendo desarrollar la interfaz de nuestras aplicaciones móviles multiplataforma una única vez, compartiendo el código de la UI. Veremos como crear aplicaciones con Xamarin.Forms además de centrarnos en cómo acceder a características propias de cada plataforma mediante la creación de servicios o Custom Renders.
¿Tienes una aplicación iOS?, ¿quieres reaprovechar tus conocimientos y código Objective-C para acceder a la plataforma universal Windows?. En esta sesión conoceremos el Bridge de Windows para iOS, convertiremos algunas aplicaciones iOS a UWP e incluso veremos como añadir características específicas de la plataforma Windows como el uso de Live Tiles por ejemplo.
Characterization of β-Glucan from Oyster Mushroom (Pleurotus pulmonarius)BRNSSPublicationHubI
The document characterizes β-glucan extracted from oyster mushrooms through alkaline extraction. β-glucan was extracted using various temperatures, potassium hydroxide concentrations, and extraction times, with the ideal parameters found to be 80°C, 30% KOH, and 90 minutes. Nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry were used to characterize the extracted β-glucan, which was found to be a β-(1-3) glucose polymer with β-(1-6) side chains and a molecular weight of 349.260 g/mol. Rheological measurements showed the β-glucan has low apparent viscosity in aqueous solutions.
Current sample preparation techniques for PFAS analysis are laborious and not easily automated. In this study, supercritical fluid extraction (SFE) was evaluated as an alternative sample preparation technique for the extraction of eighteen PFAS compounds from fish tissue, as a preconcentration step prior to their analysis by LC-MS/MS.
Effect of Ethanolic Extract of Momordica charantia on Blood Sugar Level in No...RahulGupta2015
Using four different experimental models of normal and diabetic male albino rats, blood sugar lowering efficacy of Momordica charantia Linn. of the family Cucurbitaceae has been assessed. Ethanolic (95%) extract of the whole plant of M. charantia significantly lowered blood sugar in fasted, fed and mild diabetic male albino rats at a single oral dose of 250mg/kg that has not been reported earlier. This extract also depressed the peak value significantly in the glucose loaded model.
The document summarizes the isolation and characterization of four flavone dimers - heveaflavone, amentoflavone-7",4"'-dimethyl ether, podocarpusflavone-A, and amentoflavone - from the leaves of Ouratea multiflora. The structures of the compounds were elucidated using spectral data including 2D NMR experiments. Biological assays found the isolates exhibited antibacterial activity against gram-positive bacteria and cytotoxic effects against mouse lymphoma and KB cell lines. Additionally, assays showed the extracts and isolates had antioxidant properties and some inhibited acetylcholinesterase.
Determination of Arsenic in Baby Foods and Fruit Juices by GFAASPerkinElmer, Inc.
"A complete method has been developed for the determination
of arsenic (As) in baby foods and baby fruit juices by Graphite Furnace Atomic Absorption Spectroscopy (GFAAS). This method includes sample preparation steps using microwave assisted closed vessel digestion. Foods come in a wide variety of complex sample types and matrices, but their fundamental major components are water and various carbohydrates. In this work, the samples were totally digested in a microwave oven so that the samples’ various carbohydrate matrices were completely destroyed prior to instrumental analysis."
Learn more about our solutions: http://bit.ly/1d8sGeJ
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1. DRUGS, COSMETICS, FORENSIC SCIENCES
Confirmation of Azaperone and Its Metabolically Reduced Form,
Azaperol, in Swine Liver by Gas Chromatography/Mass
Spectrometry
ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999
LAURA A. ADAM
U.S. Food and Drug Administration, Center for Veterinary Medicine, Office of Surveillance of Compliance, 7500 Standish
Pl, Rockville, MD 20855
The method described confirms the use of the
tranquilizer azaperone by detecting the parent
compound and the metabolically reduced form,
azaperol. Both are confirmed in swine liver at a tar-
get concentration of 10 ppb by gas chromatogra-
phy/mass spectrometry (GC/MS) with electron ion-
ization in the selected-ion-monitoring mode. Swine
liver tissue is ground with dry ice. Acetonitrile is
added to extract the drug from the tissue. Sodium
chloride buffer is added to the extract in prepara-
tion for solid-phase extraction (SPE). The aqueous
extract is loaded onto an SPE cartridge designed
to extract acidic and neutral drug residues from bi-
ological matrixes. The cartridge is washed with
methanol and conditioned with sodium phosphate
buffer. Azaperone and azaperol residues are eluted
with a 2% ammonium hydroxide in ethyl acetate.
The extracts are evaporated to dryness under a
stream of nitrogen and reconstituted in ethyl ace-
tate for GC/MS analysis. A DB-1 analytical column
is used to separate the compounds prior to elec-
tron ionization. The parent ion, the base peak ion,
and one diagnostic fragment ion are monitored for
both compounds. The method was validated with for-
tified tissue samples containing both azaperone and
azaperol. Azaperone-incurred tissues also were ana-
lyzed, and the presence of the parent drug and the
metabolically reduced form, azaperol, was confirmed.
A
zaperone is a neuroleptic tranquilizer belonging to the
class of butyrophenones. The current literature for
azaperone is limited, but information is available for
other related butyrophenones (1–4). The antipsychotic
butyrophenones also inhibit motor activity in animals (2).
These tranquilizers may be used therapeutically in veterinary
medicine to reduce aggressiveness and activity during live-
stock breeding (5). Azaperone is approved by the U.S. Food
and Drug Administration (FDA) for use at 2.2 mg/kg (CFR 21,
522.150) to control aggressiveness when mixing or regroup-
ing weanling or feeder pigs weighing up to 80 pounds (6).
Azaperone is not approved by the FDA for use in mar-
ket-weight swine, although it is known to be used in an ex-
tra-label manner and given prophylactically to prevent stress
in market-weight pigs during transport to the slaughterhouse.
Market-weight pigs are sensitive to stress due to transport, and
various veterinary tranquilizers are used to prevent mortality
and loss of meat quality caused by this stress. These tranquil-
izers may be administered only a few hours before slaughter
and may then give rise to residues in the animal (7, 8).
Azaperone is one of the most widely used veterinary tran-
quilizers (9). It is active at low doses (0.5 to 2.0 mg/kg), the in-
cidence of side effects is low, and it is very effective in pre-
venting traumatic shock (10). It is a short-acting drug; 16 h
after administration, it is essentially completely removed from
pig tissues (11–13).
Published analytical methods exist for azaperone and re-
lated butyrophenone tranquilizers with varying detection ca-
pabilities. In addition to residue analysis, methods have been
designed for analytical forensic toxicology and clinical chem-
istry applications. These methods use various chromato-
graphic techniques such as thin-layer chromatography
(5, 14, 15), liquid chromatography (LC; 5, 6, 16–20), and gas
chromatography (GC; 21, 22). Various mass spectrometric
(MS) techniques (23, 24) such as LC/MS (4, 16), GC/MS
(25–28), and LC-tandem MS (LC-MS/MS; 29–32) also have
been reported. However, none of these meet the Center for Vet-
erinary Medicine (CVM) requirements for confirmation of
azaperone residues.
CVM requires that a method be validated with both known
negative and fortified control samples. The method must show
specific criteria for analyte retention time matching and rela-
tive abundance matching for at least 3 diagnostic ion frag-
ments for each target. Furthermore, many of the published
procedures rely on chlorinated solvents to extract the drug
from the matrix. Use of chlorinated solvents is not desirable
for health, safety, and environmental reasons.
After our approach had been validated and found to con-
form to our criteria, azaperone-incurred tissues were gener-
ated at our facility and analyzed. The presence of both the par-
ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999 815
Received July 22, 1998. Accepted by JM January 1, 1999.
2. ent drug and the metabolically reduced form, azaperol, were
confirmed in the incurred tissue.
METHOD
Apparatus
Unless noted otherwise, equivalent apparatus and reagents
may be substituted.
(a) Centrifuge.—Beckman GPR centrifuge equipped with
a Model CH 3.7 swinging bucket rotor (Beckman/Spinco Di-
vision, Palo Alto, CA).
(b) Nitrogen evaporator.—Meyer N-EVAP analytical
evaporator, Model 111, equipped with Luer adapters for Pas-
teur pipettes (Organomation Associates, South Berlin, MA).
(c) Sample processor.—Robot Coupe sample processor,
Model RSI BX6V, equipped with stainless steel bowl and cut-
ting blades (Robot Coupe USA, Ridglan, MS).
(d) Solid-phase extraction (SPE) cartridges.—Bond Elut
Certify HF cartridges for rapid extraction of drugs of abuse,
Varian Cat. No. 1410-2081, 3 cc/300 mg (Varian, Harbor
City, CA).
(e) Vacuum manifold.—Visiprep DL SPE vacuum mani-
fold equipped with disposal flow control liners (Supelco,
Bellefonte, PA).
(f) Vortex mixer.—Vortex Genie 2, Model G-560 (Scien-
tific Industries, Bohemia, NY).
816 ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999
Figure 1. Structure and main fragment ions of azaperone.
Figure 2. Structure and main fragment ions of reduced azaperone: azaperol.
3. Reagents
(a) Ammonium hydroxide, 28.0–30.0%.—J.T. Baker
(Phillipsburg, NJ).
(b) Azaperol.—Research Diagnostics (Flanders, NJ).
(c) Azaperone.—Research Diagnostics.
(d) Hydrochloric acid (HCl), concentrated.—Fisher Sci-
entific (Fairlawn, NJ). Used to adjust pH of phosphate buffer
solution.
(e) Potassium hydroxide.—J.T. Baker. Used to adjust pH
of phosphate buffer solution.
(f) Sodium phosphate, monobasic, monohydrate, crys-
tal.—J.T. Baker.
(g) Sodium sulfate, anhydrous powder.—J.T. Baker.
(h) Solvents.—UV spectrophotometric grade ethyl ace-
tate, acetonitrile, and methanol (Burdick & Jackson,
Muskegon, MI).
(i) Stearic acid methyl ester (methyl stearate).—Sigma
Chemical (St. Louis, MO).
(j) Deionized water.—Purified through the Millipore
(Bedford, MA) Milli-Q UV plus system to a purity of
>17 MΩ/cm or equivalent. Use for all following references to
water.
(k) Dry ice pellets.—Clean pellets or chunks for grinding
tissues.
Solutions
Stability periods are noted in parentheses.
(a) Sodium chloride solution, 10% (w/v).—Store at room
temperature in a screw-capped bottle (6 months).
(b) 2% Ammonium hydroxide in ethyl acetate, elution so-
lution.—Prepare fresh daily.
(c) Phosphate buffer solution, 0.1M, pH 6.0.—Store at
ambient temperature. Inspect prior to use for any signs of con-
tamination or growth (30 days).
(d) Acetic acid 1.0M.—Store at room temperature in glass
or plastic (2 months).
(e) Azaperol stock standard, 1000 mg/mL
(RAZA-1000).—Store protected from light at 0°C or below
(6 months).
(f) Azaperone stock standard, 1000 mg/mL
(AZA-1000).—Store protected from light at 0°C or below
(6 months).
(g) AZA/RAZA mixed standard, 10 mg/mL
(AZA/RAZA-10).—Combine equal volumes of AZA-1000
and RAZA-1000. Dilute with ethyl acetate to yield a solution
containing 10 µg/mL each of AZA and RAZA. Store pro-
tected from light at 0°C or below (2 months).
(h) AZA/RAZA mixed standard, 1 mg/mL
(AZA/RAZA-1).—Dilute mixed standard AZA/RAZA-10 so-
lution to yield a solution containing 1.0 µg/mL each of AZA
and RAZA. Store protected from light at 0°C or below
(2 months). This solution is used to fortify tissue samples.
(i) Potassium hydroxide, 1.0M.—Store at ambient tem-
perature (3 months).
Animal Treatment
To generate a tissue sample containing azaperone residues
and metabolites, a male pig weighing 79 kg was intramuscu-
larly injected with 32 mg azaperone U.S.P. dissolved in etha-
nol. The animal displayed no unusual behavior after injection.
After 2 h, the pig was sacrificed, and the liver tissue was col-
lected for analysis.
ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999 817
Figure 3. Suggested fragmentation and rearrangement to yield the base peak ion at m/z 107.
4. Sample Preparation
Cut fresh livers into chunks. Pregrind dry ice to a fine pow-
der in a sample processor. Quickly drop individual liver
chunks into processor and grind into a fine powder at high
speed. Allow dry ice to sublime in a –20°C freezer, leaving a
fine powder of frozen liver.
Extraction of Samples
It is convenient to prepare 6 to 9 samples in a batch depend-
ing on positions available in the centrifuge or the evaporator.
Include at least one known negative control and one fortified
sample with each day’s test samples. Begin by weighing 10 g
portions of the uniformly ground powder. To prepare fortified
samples, add 100 µL AZA/RAZA-1. Mix on a Vortex mixer
briefly. Add 10 mL acetonitrile to each sample tube, cap the
tube tightly, and mix on a Vortex mixer for 30 s. Sonicate for
10 min. Repeat the mixing and sonication once. Centrifuge
sample at ambient temperature for 30 min at 3300 RCF (rela-
tive centrifugal force). Pour the upper acetonitrile layer into a
new tube that contains 40 mL 10% NaCl solution. Discard the
liver tissue pellet. Mix the sample tubes on a Vortex mixer.
Condition the Bond Elut Certify SPE cartridges (33) with
6 mL methanol followed by 6 mL 0.1M sodium phosphate
buffer. Transfer the aqueous extracts directly to the condi-
tioned SPE cartridge. Reduce vacuum and slowly draw the ex-
tract through the SPE until all the extract has been loaded (at
818 ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999
Figure 4. Electron ionization mass spectrum of azaperone showing the molecular ion at m/z 327 and the charac-
teristic fragment ions at m/z 233 and 107.
5. least 10 min). Rinse the charged SPE cartridges with 3 mL
1.0M acetic acid. Dry the cartridge under full vacuum for
5 min. Rinse cartridges again with 3.0 mL methanol and again
dry the cartridge. Elute with 3.0 mL ethyl acetate–ammonium
hydroxide (98 + 2) and evaporate to dryness under a stream of
nitrogen at ambient temperature. Reconstitute the dry extracts
in 50 µL ethyl acetate, briefly mix on a Vortex mixer, and
transfer to base-treated GC vials containing glass inserts to ac-
commodate the small volume. Adjust final volume to accom-
modate the sensitivity of the instrument. Inject and analyze
samples within 24 h.
Instrumental Operating Conditions
(a) GC/MS system.—Hewlett-Packard 5890 Series II gas
chromatograph equipped with a Series 5970 mass selective
detector and a Series 7673A automatic sampler
(Hewlett-Packard, Avondale, PA).
(b) Column.—DB-1 column (30 m, 0.25 µm film, and
0.25 mm od; J&W Scientific, Folsom, CA) baked at 250°C for
8 h before the daily analytical run.
(c) Injector.—Quartz 2 mm id, 250 µL, deactivated,
splitless injector liner; injector temperature, 240°C.
(d) Carrier gas.—Ultra-high-purity helium at a linear ve-
locity of 30 cm/s.
ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999 819
Figure 5. Electron ionization mass spectrum of azaperol, the reduced form of azaperone, showing the molecular
ion at m/z 329 and the characteristic fragment ions at m/z 235 and 107.
6. (e) Operating temperatures.—40°C for 1 min, rise to
140°C at 30°C/min and to 190°C at 6°C/min, maintain at
190°C for 3 min, rise to 250°C at 30°C/min, maintain for
12.3 min; total run time for each analysis, 30 min; interface
transfer line temperature, 280°C.
(f) MS analysis.—Obtain electron ionization (EI) spectra
of analytes at 70 eV. Monitor sample extracts for ions at
m/z 329, 327, 309, 233, 235, 123, 125, and 107. Monitor ion
ratios m/z 329/107 and 235/107 for azaperol and m/z 327/107
and 233/107 for azaperone. The ion at m/z 309 corresponds to
loss of water from azaperone. This ion typically is not seen in
fresh azaperol standards.
System Suitability
Conduct these tests when first establishing the analytical
system and during evaluation, to verify system suitability. Ac-
ceptable criteria for actual assays follow:
(a) Ethyl acetate blanks.—Inject a rinse of ethyl acetate to
verify baseline stability at the start of each analytical run and
after each sample.
(b) Method check.—To establish that reagents and other
aspects of the laboratory procedure are performing within ac-
ceptable limits, calculate the signal-to-noise (S/N) ratio with
the 1.0 ng/µL standard. The minimum peak-to-peak S/N ratio
must be greater than 3.
(c) Resolution and tailing.—Calculate per the current
method (34) based on the 1.0 g/µL mixed standard containing
AZA and RAZA. Resolution between the 2 peaks should be
greater than 2.0, and the tailing factors for both peaks should
be 1.2 or less. When system suitability criteria have been met,
begin analysis sequence with standards containing
1.0–4.0 g/µL azaperone and azaperol to verify the daily instru-
ment performance. Inject a solvent rinse before analysis of
known controls and before analysis of suspected samples. An-
alyze fortified samples last to avoid analyte carryover in the
820 ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999
Table 1. Validation of azaperol in swine liver tissue
fortified at 10 ppba
Replicate No.
Abundance, %
m/z 329 m/z 235 m/z 107
Day 1
1 7.8 20.0 100
2 8.9 24.1 100
Averaged standards 9.32 7.3 100
Daily limits ≤19.3 17.3–37.3 Base peak
Day 2
3 15.3 26.1 100
4 8.1 24.5 100
5 11.3 25.5 100
6 19.9 21.8 100
Averaged standards 10.0 22.9 100
Daily limits ≤20.0 12.9–32.9 Base peak
Day 3
7 19.6 9.1 100
8 15.2 26.5 100
9 13.0 23.5 100
10 10.4 27.0 100
Averaged standards 9.8 17.6 100
Daily limits ≤19.8 7.6–27.6 Base peak
a
The columns correspond to the relative abundance percentages at
each of the diagnostic ion ratios. The abundance matching limits
were determined from the averaged daily standards that
correspond to the data presented here. Any apparent outliers are
due to differences between the daily abundance limits and the
averaged abundance limits for the 3-day period.
Table 2. Validation of azaperone in swine liver tissue
fortified at 10 ppba
Replicate No.
Abundance, %
m/z 327 m/z 309 m/z 233 m/z 107
Day 1
1 5.9 8.5 22.0 100
2 6.8 9.0 25.3 100
3 5.5 8.1 21.2 100
Averaged standards 7.8 11.3 26.7 100
Daily limits ≤17.8 1.3–21.3 16.7–36.7 Base peak
Day 2
4 6.0 9.8 25.8 100
5 6.9 11.4 28.2 100
6 5.7 9.5 22.7 100
7 6.9 9.7 25.8 100
Averaged standards 7.4 12.7 24.9 100
Daily limits ≤17.4 2.7–22.7 4.9–34.9 Base peak
Day 3
8 7.5 11.9 31.2 100
9 5.2 12.5 38.2 100
10 6.9 10.6 31.5 100
Averaged standards 5.4 8.9 21.9 100
Daily limits ≤15.4 ≤18.9 11.9–31.9 Base peak
a
The columns correspond to the relative abundance percentages at
each of the diagnostic ion ratios. The abundance matching limits
were determined from the averaged daily standards that
correspond to the data presented here. Any apparent outliers are
due to differences between the daily abundance limits and the
averaged abundance limits for the 3-day period.
7. GC inlet. Reanalyze standards at the end of the run, as the GC
column is sensitive to column loading. The second set of stan-
dard injections may show greater detector response because of
carryover and column loading. Inject 1 L of the extracts onto
the column.
Results and Discussion
Rapid extraction is achieved by Vortex mixing and
sonication of samples in acetonitrile. To eliminate a
time-consuming evaporation step, the acetonitrile extract is
diluted in concentrated salt solution and applied to the SPE.
The Bond Elut Certify SPE, which is marketed for routine
testing of drugs, contains a mixed sorbent bed designed to reli-
ably extract drug residues from complex biological matrixes.
It has been used successfully, for example, to extract
haloperidol from serum or urine. It performs well with our
mixed extract, which contains high concentrations of salt and
acetonitrile, giving a clean final extract.
This method was validated by analyzing control samples
fortified with azaperone and azaperol at 10 ppb. Figures 1–3
show the possible fragmentation and structures of the diagnos-
tic ions of azaperone and azaperol. Figures 4 and 5 are the EI
mass spectra of azaperone and azaperol, respectively. With
clean control tissue obtained from a U.S. Department of Agri-
culture market pig, fortified samples were prepared at the tar-
ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999 821
Figure 6. Chromatogram of extract from a known incurred swine liver sample containing confirmed azaperone.
8. get concentration of 10 ppb. All fortified samples were con-
firmed, and all of the companion negative control samples
failed to confirm.
For a sample to be confirmed, the retention times must fall
within 10% of the acceptable retention time range of each
standard, based on the average of the standards included with
each batch. The ion ratios of a sample must match the corre-
sponding average ratios of the standards included in the analy-
sis batch within 10% absolute. Finally, the presence of both
the parent drug azaperone and the metabolically reduced form
must be confirmed to distinguish between actual misuse of
azaperone and cases of carryover from the GC inlet.
Tables 1 and 2 list ion abundances from the selec-
tive-ion-monitoring (SIM) analysis of the fortified samples.
Method performance was validated with incurred tissues from
an azaperone-dosed pig. Tissues were tested immediately af-
822 ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999
Figure 7. Chromatogram of extract from a known incurred swine liver sample containing confirmed azaperol.
9. ter slaughter and after freezing at –80°C for 6 weeks. In all
cases, azaperone use was confirmed. Figures 6 and 7 show
chromatograms of extracts from the same sample of known
incurred liver tissue that confirmed for both azaperone and
azaperol. Tables 3 and 4 list ion abundances from the SIM
analysis of fresh and frozen incurred tissues.
Conclusions
The method reliably confirmed presence of azaperone in
swine liver tissue by identifying both the parent drug com-
pound and the metabolically reduced target compound,
azaperol. By using SPE cartridges designed for drug-of-abuse
testing, we were able to keep costs down and ensure specific-
ity toward the target compound. To save time and to reduce
the number of transfer steps, the organic sample extracts were
dissolved in a large volume of salt solution. This step pre-
cluded the need for a long evaporation step and reduced sam-
ple losses due to transfer. By relying on common instrumenta-
tion to detect 2 compounds simultaneously, the method is well
suited for confirming cases of suspected extra label use of
azaperone because it reliably identifies both the parent drug
and the metabolite, azaperol, at a concentration of 10 ppb.
Acknowledgments
I thank David Heller of the Office of Research (FDA) for
discussions on mass spectrometry and Mark Henderson of the
Office of Research for generating the incurred liver tissues.
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ADAM: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999 823
Table 3. Validation of azaperol in incurred swine liver
tissuea
Replicate No.
Abundance, %
m/z 329 m/z 235 m/z 107
Day 1
1 5.3 14.3 100
2 5.8 12.9 100
3 4.9 13.5 100
4 3.4 15.2 100
5 4.6 9.3 100
6 6.0 15.0 100
Averaged standards 5.4 12.5 100
Daily limits ≤15.4 2.5–22.5 Base peak
Day 2
7 5.7 13.5 100
8 6.0 13.6 100
9 6.5 15.4 100
10 3.6 14.2 100
11 6.0 14.3 100
Averaged standards 3.5 11.8 100
Daily limits ≤13.5 1.8–21.8 Base peak
Day 3
12 1.0 8.9 100
13 4.6 11.6 100
14 4.1 17.1 100
15 2.1 14.9 100
Averaged standards 3.8 12.6 100
Daily limits ≤13.8 2.6–22.6 Base peak
a
The columns correspond to the relative abundance percentages at
each of the diagnostic ion ratios. Abundance matching limits were
calculated by averaging the daily responses. Data for day 1 were
collected from fresh tissue samples. The samples had been frozen
prior to analysis for days 2 and 3.
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